Objective To investigate the development and characterizations of the hepatocytes isolated from fetal ovine and to determine the effect of hypoxia on their growth and metabolism.Methods Fresh hepatocytes were isolated...Objective To investigate the development and characterizations of the hepatocytes isolated from fetal ovine and to determine the effect of hypoxia on their growth and metabolism.Methods Fresh hepatocytes were isolated from the liver of fetal ovine at late gestation, cultured in specific media, and exposed to normoxia(21% O2) or hypoxia(2% O2).The cellular characteristics and population purity were identified by immunocytochemistry and flow cytometry(FCM).The effects of hypoxia on cell cycle and apoptosis of the hepatocytes were evaluated by FCM, whereas the cellular ultrastructure changes were examined with a transmission electron microscope.Results The cell purity of hepatocytes was over 95%.Under hypoxia exposure, the hepatocytes showed a gradual increase in proportion at the S phase and in proliferative index, followed with a compatible increase in apoptosis and progressively decreased cell viability.Additionally, the organelles of the hepatocytes demonstrated dramatic changes, including swelling of mitochondria, disorder in cristae arrangement, expansion of endoplasmic reticulum, and a large number of circular lipid droplets emerging in the cytoplasm.Conclusion Fetal ovine hepatocytes could be primarily cultured in a short-term culture system with a high purity of over 95% and with their preserved original characteristics.Hypoxia could induce changes in ultrastructural and inhibit the proliferation of cultured fetal ovine hepatocytes through apoptotic mechanisms.展开更多
Adverse environmental factors or prenatal insults may cause developmental and pathological changes in the brain[1].Previous studies showed that a number of unfavorable prenatal factors might trigger the pathological c...Adverse environmental factors or prenatal insults may cause developmental and pathological changes in the brain[1].Previous studies showed that a number of unfavorable prenatal factors might trigger the pathological changes of adult diseases such as hypertension and diabetes mellitus[2].It is rational that inner-utero hypoxia could be a common problem of prenatal insults-induced development.展开更多
基金funded by National Natural Science Foundation [81370719 and 81671535]Jiangsu Science Foundation [BE2015642]+3 种基金Jiangsu Key Discipline of Human Assisted Reproduction Medicine Foundation [FXK201749]Jiangsu Provincial Medical Talent of the Project of Invigorating Healthcare through Science,Technology and Education [ZDRCA2016044]and Chinese Medical Association Clinical Medicine Research Reproductive Medicine [17020270696]The Priority Academic Program Development of the Jiangsu Higher Education Institutes(PAPD)
文摘Objective To investigate the development and characterizations of the hepatocytes isolated from fetal ovine and to determine the effect of hypoxia on their growth and metabolism.Methods Fresh hepatocytes were isolated from the liver of fetal ovine at late gestation, cultured in specific media, and exposed to normoxia(21% O2) or hypoxia(2% O2).The cellular characteristics and population purity were identified by immunocytochemistry and flow cytometry(FCM).The effects of hypoxia on cell cycle and apoptosis of the hepatocytes were evaluated by FCM, whereas the cellular ultrastructure changes were examined with a transmission electron microscope.Results The cell purity of hepatocytes was over 95%.Under hypoxia exposure, the hepatocytes showed a gradual increase in proportion at the S phase and in proliferative index, followed with a compatible increase in apoptosis and progressively decreased cell viability.Additionally, the organelles of the hepatocytes demonstrated dramatic changes, including swelling of mitochondria, disorder in cristae arrangement, expansion of endoplasmic reticulum, and a large number of circular lipid droplets emerging in the cytoplasm.Conclusion Fetal ovine hepatocytes could be primarily cultured in a short-term culture system with a high purity of over 95% and with their preserved original characteristics.Hypoxia could induce changes in ultrastructural and inhibit the proliferation of cultured fetal ovine hepatocytes through apoptotic mechanisms.
基金funded by National Key Support Projects[2013BAI04B05]National Natural Science Foundation[81370719 and 81671535]Jiangsu Key Discipline of Human Assisted Reproduction Medicine Foundation[FXK201749]。
文摘Adverse environmental factors or prenatal insults may cause developmental and pathological changes in the brain[1].Previous studies showed that a number of unfavorable prenatal factors might trigger the pathological changes of adult diseases such as hypertension and diabetes mellitus[2].It is rational that inner-utero hypoxia could be a common problem of prenatal insults-induced development.